| 摘要: |
| 【目的】探讨高纯度碱提灰树花菌株GF-932发酵菌丝体β-葡聚糖的免疫活性，以期为灰树花菌的开发利用提供依据。【方法】以碱提、酶解的方法，从灰树花菌株GF 932发酵菌丝体中提取β-葡聚糖，经过超滤和Q Sepharose FF 凝胶柱层析纯化冻干得高纯度β-葡聚糖，采用分子排阻法测定该β-葡聚糖的相对分子质量和分子质量分布宽度，以HPLC法测定其β-葡聚糖含量；根据人体口服推荐剂量每kg体质量2.5 mg/d，分别设低剂量组（12.5 mg/d）、中剂量组（25.0 mg/d）、高剂量组（75.0 mg/d），对SPF小鼠连续经口灌胃纯化的β-葡聚糖30 d，以灌胃灭菌蒸馏水作对照，通过迟发型变态反应、碳粒廓清和抗体生成细胞检测试验，观测灰树花β-葡聚糖对SPF小鼠吞噬细胞的足跖肿胀度、吞噬指数、溶血空斑数及小鼠体质量和脏器指数的影响。【结果】每100 g灰树花菌丝体中可得0.89 g β-葡聚糖纯品，其相对分子质量为136.43 ku，分子质量分布宽度为1.64。经口给予SPF小鼠不同剂量的灰树花β-葡聚糖30 d后，与对照组相比，各组动物体质量均呈增长状态，但差异不显著（P>0.05）。高纯度碱提灰树花β-葡聚糖能提高SPF小鼠吞噬细胞的足跖肿胀度、吞噬指数、抗体生成细胞数和脏器指数，即能增强小鼠吞噬细胞的吞噬功能及细胞、体液和免疫器官的免疫功能。【结论】高纯度碱提灰树花β-葡聚糖能显著提高小鼠的细胞吞噬功能及特异性免疫和非特异性免疫功能。 |
| 关键词: 灰树花 菌丝体 β-葡聚糖 免疫活性 |
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| 基金项目:国家重点新产品计划项目(2007GRC60031)；山东省自然科学基金项目（ZR2009DL002）;山东省高等学校科技计划项目（J09LC55） |
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| Study on immunology activities of high purified β-glucan alkali extracted from fermented mycelia of Grifola frondosa |
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| Abstract: |
| 【Objective】The immunology activities of highly purified β-glucan extracted with alkali from fermented mycelia of Grifola frondosa strain GF-932 were tested.【Method】The mycelia of G.frondosa was extracted with alkali and the crude polysaccharides were hydrolyzed with enzyme.Through the ultrafiltration,Sepharose column chromatography and lyophilization,the highly purified β-glucan was obtained.Size exclusion chromatography,liquid chromatography and software were used to determine and calculate relative molecular weight (RMW) and molecular weight distribution width of the β-glucan from mycelium,and the HPLC to assay the content of glucan.According to the recommended dose 2.5 mg/d of body mass (kg) per day in human,the G.frondosa β-glucan was formulated as a solution with sterile distilled water and administered to Specific Pathogen Free(SPF) mice at low-dose group (12.5 mg/d),middle dose group (25.0 mg/d）,high-dose group(75.0 mg/d) by gavage for 30 days.Control animals received equal volumes of sterile distilled water via the same administration.The influences of the β-glucan on the phagocytic indices phagocyte,the degree of paw swelling and the hemolysi splaque numbers by the test of clearance rate of carbon particles,delayed hypersensitivity test,and the antibody-producing cells assay was carried out respectively.The body mass and viscera index of mice was also measured.【Result】The RMW and molecular weight distribution width of the high purified β-glucan was 136.43 ku and 1.64 respectively.Gavaged at different doses of β-glucan for 30 days,the body mass of mice in every group increased,but there were no differences (P＞0.05) between any of the treatment groups and the control.The highly purified alkali extracted β-glucan from G.frondosa can enhance the phagocytic indices of phagocyte,the degree of paw swelling and the hemolysi splaque numbers of mice,and increase the viscera index in mice.This indicates that the β-glucan has the function of raising the phagocytic ability of phagocyte,the function of cellular immunity and humoral immunity,and increasing the immunologic function of immuneorgan.【Conclusion】The highly purified alkali extracted G.frondosa β-glucan can enhance evidently the immunocompetence of mice,including the phagocytic ability of phagocyte,the functions of the specifc immunity and the nonspecific immunity. |
| Key words: Grifola frondosa fermented mycelium β-glucan immunocompetence |
